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为发展国产化肾综合征出血热（ＨＦＲＳ）病毒基因工程疫苗，选择了汉滩病毒Ａ９株Ｍ基因片段为目的基因，构建转染质粒ｐＪＳＢＡ９Ｍ。以携带Ｌａｃ基因的重组病毒为亲本，使表达载体ｐＪＳＢ－Ａ９Ｍ上的Ｍ片段与痘苗病毒内的Ｌａｃ基因重组，将Ｌａｃ置换成Ａ９Ｍ片段。用蓝白斑法筛选重组痘苗病毒，经ＰＣＲ扩增证实Ａ９Ｍ片段重组入痘苗病毒基因组内。重组痘苗病毒感染的ＶｅｒｏＥ６细胞，用抗糖蛋白单克隆抗体（ＨＣＯ２、１１Ｅ１０、３Ｄ５、１６Ｄ２）间接免疫荧光法检测，呈阳性反应；放射免疫沉淀法（ＲＩＰ）证实，表达产物可与抗Ｇ２的糖蛋白单克隆抗体出现特异性沉淀带；间接免疫荧光染色法检测表明，重组病毒免疫Ｂａｌｂ／ｃ小鼠的血清与Ａ９株感染的ＶｅｒｏＥ６细胞有较好的特异性反应（１：３２０），说明Ａ９Ｍ片段在痘苗中表达成功。这为发展出血热病毒基因工程疫苗提供了一种可能的途径
In order to develop a genetically engineered vaccine against hemorrhagic fever with renal syndrome (HFRS) virus, the M gene fragment of Hantaan virus A9 strain was selected as the target gene and the transfection plasmid pJSBA9M was constructed. Using the recombinant virus carrying Lac gene as the parent, the M fragment on the expression vector pJSB-A9M was recombined with the Lac gene in vaccinia virus, and the Lac was replaced by the A9M fragment. Recombinant vaccinia virus was screened by blue-white blotting. Recombinant A9M fragment was confirmed by PCR amplification in the vaccinia virus genome. Vero E6 cells infected with recombinant vaccinia virus were detected by indirect immunofluorescence with anti-glycoprotein monoclonal antibodies (HCO2, 11E10, 3D5, 16D2). The results of radioimmunoprecipitation (RIP) The specific immunoprecipitation bands of glycoprotein monoclonal antibodies were observed. Indirect immunofluorescence staining showed that the serum of recombinant Balb / c mice immunized with recombinant virus had a good specific response (1: 320) to VeroE6 cells infected with A9 strain A9M fragment was successfully expressed in vaccinia. This provides a possible way to develop a hemorrhagic fever virus genetically engineered vaccine